TY - JOUR

T1 - 3D Bioprinting–Flow Cytometry as Analytical Strategy for 3D Cell Structures

AU - Gretzinger, Sarah

AU - Beckert, Nicole

AU - Gleadall, Andrew

AU - Lee-Thedieck, Cornelia

AU - Hubbuch, Jürgen

N1 - Funding information: This work was supported by the Helmholtz Program “BioInterfaces in Technology and Medicine (BIFTM).” We thank Professor Hartwig and co-workers from the Food Chemistry and Toxicology Group and Professor Franzreb and co-workers from the Institute of Functional Interfaces of the Karlsruhe Institute of Technology (KIT) for sharing their labs and equipment. We would also like to thank Professor Maechler from the Department of Cell Physiology and Metabolism of the University of Geneva Medical Centre, Switzerland, for providing the rat beta-cell line INS-1E. Finally, we acknowledge Stefanie Limbrunner (KIT) and Saskia Kraus (KIT) for excellent technical assistance.

PY - 2018/9

Y1 - 2018/9

N2 - The importance of 3D printing technologies increased significantly over the recent years. They are considered to have a huge impact in regenerative medicine and tissue engineering, since 3D bioprinting enables the production of cell-laden 3D scaffolds. Transition from academic research to pharmaceutical industry or clinical applications, however, is highly dependent on developing a robust and well-known process, while maintaining critical cell characteristics. Hence, a directed and systematic approach to 3D bioprinting process development is required, which also allows for the monitoring of these cell characteristics. This work presents the development of a flow cytometry-based analytical strategy as a tool for 3D bioprinting research. The development was based on a model process using a commercially available alginate-based bioink, the β-cell line INS-1E, and direct dispensing as 3D bioprinting method. We demonstrated that this set-up enabled viability and proliferation analysis. Additionally, use of an automated sampler facilitated high-throughput screenings. Finally, we showed that each process step, e.g. suspension of cells in bioink or 3D printing, cross-linking of the alginate scaffold after printing, has a crucial impact on INS-1E viability. This reflects the importance of process optimization in 3D bioprinting and the usefulness of the flow cytometry-based analytical strategy described here. The presented strategy has a great potential as a cell characterisation tool for 3D bioprinting and may contribute to a more directed process development.

AB - The importance of 3D printing technologies increased significantly over the recent years. They are considered to have a huge impact in regenerative medicine and tissue engineering, since 3D bioprinting enables the production of cell-laden 3D scaffolds. Transition from academic research to pharmaceutical industry or clinical applications, however, is highly dependent on developing a robust and well-known process, while maintaining critical cell characteristics. Hence, a directed and systematic approach to 3D bioprinting process development is required, which also allows for the monitoring of these cell characteristics. This work presents the development of a flow cytometry-based analytical strategy as a tool for 3D bioprinting research. The development was based on a model process using a commercially available alginate-based bioink, the β-cell line INS-1E, and direct dispensing as 3D bioprinting method. We demonstrated that this set-up enabled viability and proliferation analysis. Additionally, use of an automated sampler facilitated high-throughput screenings. Finally, we showed that each process step, e.g. suspension of cells in bioink or 3D printing, cross-linking of the alginate scaffold after printing, has a crucial impact on INS-1E viability. This reflects the importance of process optimization in 3D bioprinting and the usefulness of the flow cytometry-based analytical strategy described here. The presented strategy has a great potential as a cell characterisation tool for 3D bioprinting and may contribute to a more directed process development.

KW - 3D bioprinting

KW - Cell proliferation

KW - Cell viability

KW - Flow cytometry

KW - Process development

UR - http://www.scopus.com/inward/record.url?scp=85054444889&partnerID=8YFLogxK

U2 - 10.1016/j.bprint.2018.e00023

DO - 10.1016/j.bprint.2018.e00023

M3 - Article

VL - 11

JO - Bioprinting

JF - Bioprinting

M1 - e00023

ER -